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Autonomously Bioluminescent Mammalian Cells for Continuous and Real-time Monitoring of Cytotoxicity
Published on: October 28, 2013
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A novel thermostable beetle luciferase based cytotoxicity assay
Sunju Choi1, Hittu Matta1, Ramakrishnan Gopalakrishnan1
1Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
Scientific Reports
|May 12, 2021
Summary
A new Matador-Glo assay uses a thermostable Click Beetle Luciferase for sensitive cancer immunotherapy testing. This cost-efficient method aids in cell death detection and in vivo imaging applications.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Cytotoxicity assays are crucial for developing cancer immunotherapies.
- Previous work introduced the marine luciferase-based Matador assay.
- A need exists for improved, efficient cell death detection methods.
Purpose of the Study:
- To develop a novel, sensitive cytotoxicity assay named Matador-Glo.
- To utilize a thermostable Click Beetle Luciferase variant (Luc146-1H2) for luminescence detection.
- To establish a versatile tool for cell death detection and in vivo imaging.
Main Methods:
- Ectopic overexpression of Luc146-1H2 in target cells.
- Luminescence detection using Luc146-1H2 and D-luciferin substrate.
- Assessment of membrane integrity loss in dead/dying cells.
- Combination with the marine luciferase-based Matador assay for dual detection.
Main Results:
- The Matador-Glo assay is simple, fast, specific, sensitive, and cost-efficient.
- Luc146-1H2 activity is measurable in dead and dying cells upon membrane damage.
- The assay can be combined with the Matador assay for dual luciferase cell death detection.
- Luc146-1H2 expressing cells are suitable for in vivo bioluminescence imaging.
Conclusions:
- The Matador-Glo assay offers a robust and efficient method for cytotoxicity testing.
- This assay provides a valuable tool for cancer immunotherapy development and cell death research.
- The technology supports both in vitro and in vivo applications, enhancing its utility.
